Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (<i>Artocarpus </i><i>champeden</i>) vinegar prepared with maltodextrin and chitosan as wall materials

Anton Rahmadi; Siti Nurjannah; Yulian Andriyani; Maghfirotin Marta Banin; Miftakhur Rohmah; Nur Amaliah; Kartika Sari; Aswita Emmawati

doi:10.12688/f1000research.109612.1

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Research Article

Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (Artocarpus champeden) vinegar prepared with maltodextrin and chitosan as wall materials

[version 1; peer review: 1 approved with reservations]

Anton Rahmadi^1,2, Siti Nurjannah¹, Yulian Andriyani¹, [...] Maghfirotin Marta Banin¹, Miftakhur Rohmah¹, Nur Amaliah¹, Kartika Sari^1,3, Aswita Emmawati¹

Anton Rahmadi^1,2, Siti Nurjannah¹, [...] Yulian Andriyani¹, Maghfirotin Marta Banin¹, Miftakhur Rohmah¹, Nur Amaliah¹, Kartika Sari^1,3, Aswita Emmawati¹

PUBLISHED 29 Jul 2022

Author details Author details

¹ Department of Agricultural Product Technology, University of Mulawarman, Samarinda, East Kalimantan, 75119, Indonesia
² Research Center for Drugs and Consmetics from Tropical Rain Forest (PUI-PT Oktal), University of Mulawarman, Samarinda, 75123, Indonesia
³ Agricultural Sciences Doctoral Program, University of Mulawarman, Samarinda, East Kalimantan, 75123, Indonesia

Anton Rahmadi
Roles: Funding Acquisition, Investigation, Project Administration, Supervision, Writing – Review & Editing

Siti Nurjannah
Roles: Formal Analysis

Yulian Andriyani
Roles: Data Curation, Validation

Maghfirotin Marta Banin
Roles: Methodology, Project Administration

Miftakhur Rohmah
Roles: Conceptualization, Investigation, Methodology, Writing – Original Draft Preparation

Nur Amaliah
Roles: Investigation, Methodology

Kartika Sari
Roles: Data Curation, Resources

Aswita Emmawati
Roles: Data Curation, Supervision, Writing – Original Draft Preparation

OPEN PEER REVIEW

REVIEWER STATUS

This article is included in the Agriculture, Food and Nutrition gateway.

Abstract

Background: Mandai cempedak vinegar (MCV) is a fermented vinegar produced from the inner skin of cempedak (Artocarpus champeden), which contains antioxidants classified as flavonoids, phenols, and tannins. These bioactive compounds are sensitive to heat and prone to oxidative damage. Therefore, an encapsulation process is proposed to protect the bioactive compounds. This study aimed to design a potential scaling-up formulation of spray-dried encapsulated MCV based on Total Soluble Solid (TSS) with the addition of maltodextrin and chitosan, followed by determining the nutrition and phytochemical values of the formulation.
Methods: The formulation employed maltodextrin to achieve TSS of 15, 20, and 25 ºBrix as the primary wall material treatment factor. The second factor was chitosan as auxiliary wall material at 1, 2, and 3% (w/w of maltodextrin). Products were spray-dried at 100 ºC inlet temperature and 80 ºC outlet temperature. Analyses of nutrition, flavonoid, phenol, and tannin were conducted in triplicate for each encapsulated product.
Results: The 15 ºBrix of TSS from maltodextrin with 1% chitosan emerged as the best-encapsulating material, giving 7.54% moisture, 0.75% ash, 0.42% protein, 0.35% fat, and 90.94% carbohydrate content, resulting in a phytochemical activity equivalent to 9.13 mg Catechin Equivalent kg^-1, 69.61 mg Gallic Acid Equivalent kg^-1, and 25.04 mg Tannic Acid Equivalent kg^-1. Compared to maltodextrin, the chitosan generally contributed less to the proximate, flavonoid, phenol, and tannin content of the encapsulated MCV.
Conclusions: The best formulation contained maltodextrin at 15 ºBrix of TSS and 1% chitosan. Maintaining optimum TSS was a key to producing consistent encapsulated MCV with high phytochemical activity.

Keywords

chitosan, maltodextrin, mandai cempedak vinegar, spray drying, total soluble solids

Corresponding author: Anton Rahmadi

Competing interests: No competing interests were disclosed.

Grant information: This work was supported by the Indonesian Ministry of Education, Culture, Research, and Technology, under grant no.: 598/UN17.L1/PG/2021, awarded to Anton Rahmadi.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Copyright: © 2022 Rahmadi A et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

How to cite: Rahmadi A, Nurjannah S, Andriyani Y et al. Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (Artocarpus champeden) vinegar prepared with maltodextrin and chitosan as wall materials [version 1; peer review: 1 approved with reservations]. F1000Research 2022, 11:865 (https://doi.org/10.12688/f1000research.109612.1) First published: 29 Jul 2022, 11:865 (https://doi.org/10.12688/f1000research.109612.1) Latest published: 29 Jul 2022, 11:865 (https://doi.org/10.12688/f1000research.109612.1)

Introduction

Mandai cempedak is a traditional food fermented from the inner skin of cempedak (Artocarpus champeden), technologically processed with the addition of a bacterial starter containing Lactic Acid Bacteria (LAB).¹ The liquid separated from mandai cempedak solids is named mandai cempedak vinegar (MCV). MCV is produced twice as much as the weight of the solid part during fermentation. MCV developed in our design is a by-product of mandai cempedak fermentation with Lactobacillus casei as a starter.¹^–³ Mariana et al.⁴ show that MCV can reduce cholesterol levels in mice after 14 days of intervention. In addition, MCV contains bioactive, i.e., lactic acid, flavonoid compounds, and saponins. These substances may reduce cholesterol levels.⁵^,⁶

MCV is easily degraded during storage because of its content. The changes include oxidation of phenol and evaporation of volatile compounds. In addition, the liquid form of MCV is affected by environmental factors, i.e., temperature, oxygen, humidity, light, and other unfavorable conditions during the storage process.⁷^,⁸ Therefore, the technology has been developed to protect the active compounds in MCV by converting MCV into encapsulated form. Encapsulation is the process of protecting, enclosing, or trapping the core material (liquid, gas, and solid particles) in a solid shell in the form of coating material for controlled release, immobilization, protection, or isolation of core material.⁹

The composition of coating materials can affect the characteristics of the resulting encapsulated product.¹⁰ This study used maltodextrin and chitosan as wall and auxiliary coating materials in producing encapsulated MCV. It is essential to maintain the stability of the Total Soluble Solid (TSS) value in the encapsulation process.¹¹ As customarily observed from natural products,¹² MCV has a tiny but inconsistent amount of soluble solids. Hence, maltodextrin contributes to TSS as a primary controlling factor for wall material.¹³ Maltodextrin is used because it has a high solubility in water, high viscosity, low price, and can maintain the stability of polyphenol compounds in a formulation.¹⁴^,¹⁵

The auxiliary coating material used in this study is chitosan. Chitosan is non-toxic and biodegradable with some antimicrobial properties, benefiting product shelf life extension.¹⁶^,¹⁷ In addition, chitosan is a biocompatible amino polysaccharide capable of forming ionic or covalent bonds with crosslinking agents and forming networks where the active compounds can be retained.¹⁸ These properties are helpful for encapsulation material of various active ingredients, enveloping bioactive compounds, and reducing oxidation.¹⁷^,¹⁹ As a result, chitosan is widely used as an encapsulation agent for sensitive core materials in the pharmaceutical industry, such as lipophilic drugs,²⁰ vitamin D2,²¹ ampicillin,²² and olive oil extract.²³ However, chitosan is expensive. Therefore, it is used as auxiliary material in this research.

Spray drying may be applied to encapsulate bioactive material from MCV because the short contact time with the medium and the high evaporation rate results in a high-quality product compared to conventional drying methods.²⁴ As a result, the spray drying technique is relatively easy to apply in the scaling-up process for volatile or non-volatile oils and flavor encapsulation. In addition, the production cost is relatively low compared to other available technologies.²⁵^,²⁶

This research aims to design a potential scaling-up formulation of spray-dried encapsulated MCV based on maltodextrin equivalent to specified TSS with the addition of chitosan. The experimental parameters include proximate values (moisture, ash, protein, fat, and carbohydrate contents) and phytochemical characteristics (flavonoid, tannin, and phenol). The encapsulation of MCV is expected to be a new niche seasoning flavor with the additional advantage of high bioactive capacity.

Methods

Mandai Cempedak Vinegar (MCV)

The process of making MCV is described by Mariana et al.⁴ The cempedak was obtained from the traditional markets in Samarinda, East Kalimantan, Indonesia. The inner cempedak skin was cut into small pieces, cleaned with running water, and then boiled with a ratio of 1:2 (w/v) cempedak and water at 100°C for five minutes to remove the sap. The cempedak skin was then cooled to achieve room temperature (28±3°C) in a closed jar. The boiling procedure was repeated twice. The boiled cempedak skin was then placed in a sterile container, and boiled potable water was added (2:1 v/w). LAB starter was added at 4% (v/v) and was aseptically stirred until evenly distributed. The mandai cempedak was fermented for 14 days at 5-8°C (stored in the refrigerator). Subsequently, the mandai cempedak was crushed with a blender and filtered with a clean cloth. The filtered MCV was then centrifuged at 3000 revolutions per minute (RPM) for 15 minutes. The supernatant was collected and stored at 5-8°C.

MCV encapsulation

MCV encapsulation was carried out using a spray dryer (locally built by Maxer Sterile - Malang, Indonesia, with a capacity of 0.5-5 liter/hour, electric heating of 13 kilowatts, automatic temperature control, a yield of 8-12%, and moisture content of 3-7%). Each batch consisted of 500 mL MCV. The treatment used in the encapsulation of MCV was maltodextrin equivalent to TSS 15, 20, 25 °Brix and chitosan (1, 2, 3% w/w of maltodextrin). About 500 mL MCV and a specified maltodextrin content by TSS value were homogenized. The intended Brix value was achieved by adding maltodextrin in a stepwise manner. TSS was measured by a hand refractometer (Brix scale: 0 to 32% Brix, Automatic Temperature Compensation Range: 10-30°C) at room temperature of 28±3°C. The range of maltodextrin addition was 70-170 g, corresponding to 15 – 25 °Brix. Chitosan was added according to the treatment with concentrations of 1, 2, and 3% by weight of the maltodextrin used. The mixtures were spray-dried with the condition of 100°C inlet temperature and 80°C outlet temperature. The obtained encapsulated MCV was stored at -20±3°C for further analysis.

Moisture content analysis

The water content analysis was carried out by referring to the modified Lindani²⁷ method regarding the number of samples and the time of sample testing. A total of 0.5 grams of sample was put into the moisture analyzer (OHAUS MB 125), then the sample was flattened on a pan, and the instrument was closed again. The moisture was measured as per recommended method by the manufacturer. The results obtained were then recorded.

Ash content analysis

The measurement of ash content used the Andarwulan et al.²⁸ method. The inorganic residue formed by burning off the organic matter of the sample in a muffle furnace at 550±50°C for four hours was referred to as ash. A pre-heated crucible was filled with 2 g of the sample. The crucible was placed in a muffle furnace at 550±50°C for four hours or until whitish-grey ash was formed. After that, the crucible was placed in the desiccator and weighed.

Protein content analysis

Crude protein is calculated by multiplying the nitrogen content of the food by a factor of 6.25. This number is used because most protein contains 16% nitrogen. The Kjeldahl method is used to determine crude protein.²⁹ Digestion, distillation, and titration are all part of the process.³⁰ About 2 g of the material was weighed carefully in a Kjeldahl flask. About 25 ml of 97% sulfuric acid to the flask and 1 g of Kjeldahl tablet (containing Na₂SO₄ 96.5%, CuSO₄ 1.5%, Se 2.0%) (Merck, USA) were added. Heat was slowly applied in a fume cupboard to avoid excessive frothing, then digestion continued for 45 minutes, or until the digesta became transparent pale green. The digesta was allowed to cool before adding 100 ml of distilled water. The digesta was rinsed 2-3 times before adding the digesting flask to the bulk. Distillation: Markham distillation device was used for distillation. The digesta was taken 10 ml of the digest to boil in the distillation equipment after heating it. About 10 ml sodium hydroxide (10 mol/l) (Merck, USA) was added to avoid ammonia loss. Five drops of methyl red indicator (0,1 % in ethanol) (Sigma-Aldrich, USA) were distill-filtered into 50 ml of 2% boric acid (Merck, USA). Titration: The alkaline ammonium borate was directly titrated with 0.1N HCl (Merck, USA). The volume of acid used was recorded as the titer value. The amount of acid used was factored into the formula, yielding %N.

Total - N = \frac{mL NaOH (sample titration - blank titration) \times N NaOH \times 14,008 \times 100}{sample weight (g) \times 100}

Percent protein = (Total - N) \times 6, 25

Constant: Nitrogen molecular weight 14,008 g/mol; Conversion factor: 6,25

Fat content analysis

Crude lipid was determined using the Soxhlet extraction technique (Association of Official Analytical Chemists, 2005).³¹ First, the lipid content of the sample (2 g) was extracted using 100 mL of petroleum ether. Then, the mixture was filtered, and its lipid content was collected in a pre-weighed (W₁) clean beaker. After that, exhaustive lipid extraction was done on the same sample with 100 mL of n-hexane (Fulltime, China) for 24 h. It was then filtered and decanted into a beaker (W₁). Next, the lipid content was concentrated to dryness in a steam bath and oven-dried at 40–60 °C, and the beaker was reweighed (W₂). Finally, the percentage of lipid was calculated.

Fat content (%) = \frac{(W_{3} - W_{2})}{W_{1}} \times 100 %

Where:

W₁=Weight of sample (g)

W₂=Weight of fat flask without fat (g)

W₃=Weight of fat flask with fat (g)

Carbohydrate content by difference

The carbohydrate content was calculated by subtracting the total crude protein, ash, and crude fat from the total dry matter.³²

Carbohydrate (%) = 100 % - (% Moisture + % Ash + % crude fat + % crude protein)

Total flavonoid

The total flavonoid content was determined.³³ Encapsulated MCV was weighed at 1 mg and then dissolved in 10 mL of 95% ethanol (SmartLab, Indonesia). A total of 0.7 mL of distilled water was added to the encapsulation and dissolved in ethanol. Then, about 0.1 mL of 5% NaNO₂ (Sigma-Aldrich, USA) was added to the mixture. After five minutes, 0.1 mL of 10% AlCl₃ (Sigma-Aldrich, USA) was added. After six minutes, 0.5 mL of 1 M NaOH (Merck, USA) was added. All ingredients were mixed with a vortex evenly, then incubated for 10 minutes. The absorbance value was detected at a wavelength of 510 nm (Biospectrometer, Eppendorf, Germany). The results obtained were plotted against the catechin (Sigma-Aldrich, USA) standard curve prepared similarly. Total flavonoid was expressed as milligrams of catechin equivalent per kilogram dry weight extract.

Total phenol

Measurement of total phenol was conducted.³⁴^,³⁵ About 0.3 g encapsulated MCV was carefully dissolved in 10 mL of ethanol and distilled water (1:1 ratio). About 0.2 mL of the extract solution was pipetted and added with 15.8 mL of distilled water and 1 mL of Folin-Ciocalteu 50% (v/v) reagent (Merck, USA). The solution was allowed to stand for eight minutes, and then 3 mL of 5% (w/v) Na₂CO₃ (Sigma-Aldrich, USA) was added. Then the solution was allowed to stand for two hours in the dark at room temperature (28±3°C). The absorbance value was detected at a wavelength of 725 nm (BioSpectrometer, Eppendorf, Germany). The absorbance value was plotted against a gallic acid (Sigma-Aldrich, USA) standard curve prepared similarly. Total phenol was expressed in milligrams of gallic acid equivalent per kilogram dry weight extract.

Total tannin

The total tannin was calculated.³⁶ A total of 0.5 g of encapsulated MCV was macerated in 10 mL of diethyl ether (Merck, USA) for 20 hours, then filtered, and the residue obtained was boiled with 100 mL of distilled water for two hours, then allowed to cool. About 0.1 mL of the cold sample was added with 0.1 mL of Folin-Ciocalteu reagent (Merck, USA) and vortexed, added with 2 mL of 2% Na₂CO₃ (Sigma-Aldrich, USA), and vortexed again. The absorbance value was detected at a wavelength of 760 nm (BioSpectrometer, Eppendorf, Germany) after being incubated for 30 minutes at room temperature (28±2°C). The results obtained were plotted against a tannic acid (Sigma-Aldrich, USA) standard curve prepared similarly. Thus, the total tannin content was expressed in milligrams tannic acid per kilogram extract.

Statistical analysis

The experimental design used in this study was a factorial, completely randomized design with nine treatments and two replications. This study consisted of two factors, namely maltodextrin equivalent to TSS (15, 20, 25 °Brix) and chitosan (1, 2, 3% w/w maltodextrin), as shown in Table 1.

Table 1. Mandai cempedak vinegar encapsulation treatment.

Sample code	Treatment
F1	TSS 15 °Brix, K 1%
F2	TSS 15 °Brix, K 2%
F3	TSS 15 °Brix, K 3%
F4	TSS 20 °Brix, K 1%
F5	TSS 20 °Brix, K 2%
F6	TSS 20 °Brix, K 3%
F7	TSS 25 °Brix, K 1%
F8	TSS 25 °Brix, K 2%
F9	TSS 25 °Brix, K 3%

The data obtained from the chemical and phytochemical tests were analyzed using variance analysis at a 5% significance level using a GraphPad Prism (v9.2), but can also be analyzed using any open-source statistical software, including the freely available R. If the variance test shows a significant treatment effect, the variance analysis was continued with the Tukey test.

Result and discussion

Moisture content

Moisture content is an important parameter determining dry products’ quality, such as dry encapsulation. The lower moisture content makes the product protected from bacterial and fungal spoilage.³⁷ The moisture content of the MCV encapsulated product in this study ranged from 5.31 ± 1.00% to 10.45 ± 0.85% (see Table 2 and Underlying data⁶⁷). The TSS treatment and the interaction between the two factors had a significant effect (p<0.05) on the moisture content of the encapsulated MCV product. In contrast, the chitosan treatment showed no significant effect (p>0.05). However, TSS treatments showed a significantly different effect on moisture content. The 15 °Brix treatment group generally produced more moisture content than the 20 °Brix treatment group. The higher the TSS used in the encapsulation process, the lower the percentage of moisture content (Table 2).

Table 2. Proximate values of encapsulated mandai cempedak vinegar product.

Sample	Moisture (%)	Ash (%)	Protein (%)	Fat (%)	Carbohydrate (%)
F1	7.54±0.25b	0.75±0.35	0.42±0.02a	0.35±0.00a	90.94±0.16
F2	10.45±0.85c	1±0.00	0.59±0.06bc	0.38±0.00b	87.59±0.23
F3	8.02±0.04b	1±0.00	0.69±0.01e	0.40±0.00c	89.89±0.01
F4	7.15±0.58b	1.25±0.35	0.63±0.02cd	0.69±0.00d	90.29±0.24
F5	5.31±1.00a	0.75±0.35	0.70±0.05d	0.69±0.00de	92.55±0.35
F6	7.43±0.28b	1±0.71	0.46±0.04a	0.69±0.00e	90.42±0.26
F7	7.45±0.16b	0.5±0.00	0.52±0.04ab	0.81±0.00f	90.73±0.05
F8	7.70±0.14b	0.25±0.35	0.57±0.06bc	0.85±0.00g	90.63±0.14
F9	8.47±1.12b	0.5±0.71	0.46±0.05a	0.85±0.00g	89.73±0.47

According to Marte et al.,³⁸ the higher the total solids dried to a certain extent, the higher the evaporation rate, causing lesser moisture content. Hayati et al.³⁹ stated that maltodextrin used in quantities could reduce the moisture content of the encapsulated results.

At TSS 25 °Brix, the encapsulated MCV experienced an increase in moisture content. Different total solids influence the increase in moisture content. The high total solids are related to the drying rate during spray drying.⁴⁰ However, a high concentration of maltodextrin causes coagulation.⁴¹ As a result, the water molecules that will diffuse are blocked by larger maltodextrin molecules and cause a decrease in the drying rate, thus causing the moisture content to increase.⁴¹

Chitosan has no significant effect on the moisture content of the encapsulated MCV product. However, chitosan has a humidifying effect, retaining moisture during the spray drying. Moreover, chitosan can slow down the crystallization process during spray drying.⁴²

Ash content

The ash content of the MCV encapsulated product in this study ranged from 0.25±0.35% to 1.25±0.35% (see Table 2 and Underlying data⁶⁷). The treatment and interaction of TSS and chitosan had no significant effect on the ash content of the encapsulated MCV product. The highest ash content value was obtained in the TSS 20 °Brix treatment with 1% chitosan, and the lowest ash content was found in the TSS 25 °Brix treatment with 2% chitosan.

Ash content is usually pointing to the total minerals in a material. The ash content of the MCV encapsulated product showed no significant difference. The non-significance values happened because the raw material in MCV was used in the same concentration. Siregar et al.⁴³ concluded ash content of the mandai cempedak product per 100 g of dry matter was 4.3%. The low ash content obtained is because MCV is a derivative product of the mandai. Caez and Jaraba⁴⁴ reported 0.43% ash content in mango juice microencapsulated with maltodextrin compared to other encapsulated products. Minerals are not significantly affected by chemical and physical treatments during processing.⁴⁵ For example, specific mineral ions can be oxidized in the presence of oxygen, but their final concentrations were not significantly affected.⁴⁶

Protein content

The crude protein content in this study ranged from 0.42±0.02% to 0.70±0.05% (see Table 2 and Underlying data⁶⁷). The treatment of TSS and chitosan and the interaction between the two factors significantly affected the protein content of the Encapsulated MCV. The TSS and chitosan treatments have significantly different effects. The highest protein content was obtained from the TSS 20 °Brix treatment with 2% chitosan, and the lowest protein content was obtained from the TSS 15 °Brix treatment with 1% chitosan. The TSS treatment of 15 and 20 °Brix showed no significant difference, but protein levels decreased in TSS 25 °Brix. The value explains that the higher the total solids used in the Encapsulated MCV, the lower the protein content of the powder produced.

Proteins consist of long chains of amino acids bound to each other by peptide bonds. Amino acids consist of carbon, oxygen, hydrogen, and nitrogen.⁴⁷ The decrease in protein content in the processing of MCV encapsulated products is also due to the spray drying method used, which causes the particles to come into direct contact with high temperatures, which allows the denaturation process to occur. For example, Anandharamakrishnan et al.⁴⁸ reported that protein denaturation by the spray drying method could occur even at an outlet temperature of 60°C.

The higher the chitosan concentration in the processing of the encapsulated MCV, the higher the protein content of the powder produced. Commercial chitosan contains 0.48% protein, so there may be an increase in protein due to the higher amount of chitosan added.⁴⁹ Conversely, a concentration of 3% chitosan showed a decrease in protein levels. This was because chitosan concentration was too high, making chitosan insoluble. This situation causes a crack in the encapsulation wall so that the compound can diffuse out through the gap formed.⁵⁰

Fat content

Fats are compounds formed from the esterification reaction between fatty acids and glycerol. The use of high temperatures in fats will cause the double bonds in the fat to break, breaking the fat into glycerol and fatty acids.⁵¹ The crude fat content value of the encapsulated MCV in this study ranged from 0.35% to 0.85% (see Table 2 and Underlying data⁶⁷). The treatment of TSS and chitosan and the interaction between the two factors significantly affected the fat content of the encapsulated MCV. The highest fat content was obtained from the TSS 25 °Brix treatment with 3% chitosan, and the lowest fat content was obtained from the TSS 15 °Brix treatment with 1% chitosan. The fat content of the encapsulated MCV increased with the increasing amount of TSS and chitosan used.

The higher the maltodextrin and chitosan, the higher the fat content of the encapsulated MCV. Therefore, the rehydration of the sample may occur in a small amount. However, it affects the total weight of the fat tested. Maurer et al.⁵² stated that rehydration might affect the fat content measurement. Since the fat content of encapsulated MCV is less than 1%, the minute increase in fat concentration affects the statistical calculation.

The fat content in cempedak fruit is less than 2%, so the fat content produced in encapsulated MCV is also insignificant.⁵³ On the other hand, the low-fat content provides advantages for the drying process and extends the shelf life.⁵⁴^,⁵⁵

Carbohydrate content

In this study, the carbohydrate content of the encapsulated MCV was determined by the by-difference method. Major nutrients that affect carbohydrate levels include protein, fat, moisture, and ash content.⁵⁶ In this study, the carbohydrate content values ranged from 87.59±0.23% to 90.94±0.16% (see Underlying data⁶⁷). The maltodextrin equivalent to TSS and chitosan treatments and their interactions had no significant effect on the carbohydrate content of the encapsulated MCV.

The carbohydrate content of cempedak fruit is 84-87%, so the carbohydrate content dominates the encapsulated MCV.⁵³ This is also possible because of maltodextrin and chitosan’s combined carbohydrate content. Maltodextrin is a saccharide polymer generally obtained from the hydrolysis of corn starch with acids or enzymes.⁵⁵ Chitosan is a non-toxic amino polysaccharide obtained from chitin through a complete deacetylation process.¹⁸ In this study, the carbohydrate content of the MCV encapsulated product was not significantly different, only slightly influenced by the increase in wall material concentration. In another report, the carbohydrate content in the encapsulated gac fruit did not change with the addition of auxiliary material.⁵⁷

Total flavonoid

Before the spray drying, the total flavonoid content in MCV was 11.4 mg Catechin Equivalent (CE) kg^-1. The flavonoid content of encapsulated MCV varies with maltodextrin and chitosan. The total flavonoid value of the MCV encapsulated product ranged from 6.99±1.08 mg CE (Catechin Equivalent) kg^-1 to 9.13±0.89 mg CE kg^-1 (see Table 3 and Underlying data⁶⁷). The highest flavonoid content was found in encapsulated product with 15 °Brix TSS and 1% chitosan, the encapsulation with the lowest concentration of wall material. The encapsulation concentration influenced the flavonoid content in purple sweet potato in another research.⁵⁸ The binding of flavonoid by the encapsulated matrix caused the product with a higher encapsulated concentration to bind more flavonoid.⁵⁸ As a result, an increase in the concentration of the encapsulant causes a decrease in flavonoid levels.

Table 3. Phytochemical values of encapsulated mandai cempedak vinegar.

Sample	Flavonoid (mg CE kg^-1)	Phenol (mg GAE kg^-1)	Tannin (mg TAE kg^-1)
F1	9.13±0.89	69.61±1.70bc	25.04±0.17f
F2	7.38±0.09	79.52±1.57d	26.67±0.64g
F3	8.23±0.94	83.13±0.92e	19.40±0.40d
F4	8.91±1.23	72.30±2.10c	13.12±0.89a
F5	8.96±0.48	56.93±0.52a	14.63±0.65b
F6	7.56±3.13	80.81±1.83de	15.42±0.07b
F7	6.99±1.08	68.41±1.05b	15.77±0.02b
F8	7.36±2.87	81.93±1.57de	22.00±0.45e
F9	8.18±2.40	70.81±0.52bc	17.12±0.69c

The treatment of TSS and chitosan and their interactions had no significant effect on the total flavonoid of the Encapsulated MCV. The increase in maltodextrin and chitosan showed no significant difference in the total value of encapsulated flavonoid. The use of maltodextrin and chitosan can protect flavonoid compounds well because they have properties that can prevent oxidation.⁴¹^,⁵⁹

Total phenol

The initial total phenol content in MCV was 103.31 mg GAE (Gallic Acid Equivalent) kg^-1. The total phenol value of the MCV encapsulated product in this study ranged from 56.93±0.52 mg GAE kg^-1 to 83.13±0.92 mg GAE kg^-1 (see Table 3 and Underlying data⁶⁷). The treatment of maltodextrin and chitosan and their interactions had a significant effect on the total value of the encapsulated product of MCV. Each treatment of TSS and chitosan and their interactions gave significantly different effects. The highest total phenol was obtained from 15 °Brix TSS treatment with 3% chitosan, and the lowest total phenol was obtained from 20 °Brix TSS treatment with 2% chitosan.

The highest total phenol was obtained from the 15 °Brix TSS treatment, and the lowest was accepted from the 20 °Brix TSS treatment. The higher the total solids, the total encapsulated phenol tends to decrease. Safithri et al.⁶⁰ stated that the more total solids used in functional drinks based on red betel leaf extract (Piper crocatum) in maltodextrin as coating material, the lower the total coated phenol. In addition, encapsulation materials such as maltodextrin and chitosan coat bioactive compounds in the form of phenol compounds.⁶¹ When tested with phenol reagents, not all bioactive components can react, resulting in under-valued measurement.⁶² The oxidation process causes this total degradation of phenol due to heat treatment during spray drying activities.⁶³ This can be related to changes in the molecular structure that cause a decrease in reactivity. In addition, the possibility of a reduction of total phenol levels is due to the formation of phenol-protein complexes or decomposition of phenol compounds and changes in chemical structure due to pressure and temperature.⁶³

The highest total phenol was obtained from 3% chitosan treatment, and the lowest was accepted from 1% chitosan treatment. The use of higher concentrations of chitosan can encapsulate more phenolic compounds. The trend is also following the research of Kistriyani et al.,⁵⁹ which states that the higher the chitosan content, the higher the binding ability to protect phenolic compounds. Belscak-Cvitanovic et al.⁶⁴ showed that chitosan as an encapsulant was very effective in safeguarding bioactive compounds, slowing their release profile, and increasing encapsulation efficiency. Encapsulation efficiency is calculated based on the number of polyphenols encapsulated. Encapsulation efficiency is related to the particles’ morphological characteristics and heat resistance. Therefore, chitosan can protect the encapsulated active compounds.

Total tannin

The total tannin content in MCV was 33.63 mg TAE (Tannic Acid Equivalent) kg^-1. The total tannin value of the MCV encapsulated product in this study ranged from 13.12±0.89 mg TAE kg^-1 to 26.67±0.64 mg TAE kg^-1 (see Table 3 and Underlying data⁶⁷). The highest total tannin was found in 15 °Brix TSS treatment with 2% chitosan. The treatment of maltodextrin and chitosan and their interactions significantly affected the total tannin of the encapsulated MCV.

The highest total tannin was found in the 15 °Brix TSS treatment, and the lowest was obtained from the 20 °Brix TSS treatment. The increase in TSS at various concentrations of maltodextrin and chitosan causes the total tannin content to be lower. The increasing TSS affects the movement of volatile components to the dried droplet surface.⁵⁰ According to Rosenberg et al.,⁶⁵ the concentration of total solids affects the viscosity and decreases the volatile components.

The more the addition of chitosan to various TSSs, the higher the total encapsulated tannin. Therefore, the ability of chitosan to protect the tannin compounds in the product to be dried by the spray drying process is more significant. Chitosan protects essential compounds contained in materials such as bioactive components, including tannin, because chitosan has a solid binding capacity to coated bioactive components.⁶⁶ The concentration of 3% chitosan showed a decrease, and this happened because chitosan was given in large quantities so that the chitosan became insoluble. Ali et al.⁵⁰ stated that increasing the higher chitosan concentration could cause chitosan to become insoluble. This condition results in cracks in the crust so that the bioactive components diffuse out through the gaps formed.

Conclusions

Maltodextrin and chitosan treatment significantly affected moisture, protein, and fat contents. Chitosan has a humidifying effect and can slow down the crystallization process during spray drying. In addition, a high concentration of maltodextrin causes coagulation; water molecules that will diffuse are blocked by larger maltodextrin molecules and cause a decrease in the drying rate, thus causing the moisture content to increase. The decreased protein content is due to the spray drying method used. However, the higher the chitosan concentration, the higher the protein content of the powder produced because commercial chitosan contains 0.48% protein, the more chitosan used, the higher the protein content. On the other hand, the higher the maltodextrin and chitosan, the higher the fat content, and the rehydration of the sample may occur in a small amount. Total flavonoid, phenol, and total tannin were related to maltodextrin equivalent to TSS and chitosan treatment. The best phytochemical concentrations of encapsulated MCV were produced from 15 °Brix TSS with 1% chitosan. Encapsulated MCV product with this treatment has 7.54% moisture, 0.75% ash, 0.42% protein, 0.35% fat, 90.94% carbohydrate contents, giving bioactive capacities equivalent to 9.13 mg CE kg^-1, 69.61 mg GAE kg^-1, and 25.04 mg TAE kg^-1.

Data availability

Underlying data

Open Science Framework: MCV encapsulation treatment. https://doi.org/10.17605/OSF.IO/3AQSW.⁶⁷

This project contains the following underlying data:

▪ MCV encapsulation treatment.csv (sample code/key and treatment).
▪ Ash content.csv (replication and average ash on MCV encapsulation).
▪ Ash_data for graphs with GraphPad.csv (ash percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Ash_Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Ash_Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).
▪ Carbohydrate Content.csv (replication and average carbohydrate on MCV encapsulation).
▪ Carbohydrate _data for graphs with GraphPad.csv (carbohydrate percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Carbohydrate _Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Carbohydrate _Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).
▪ Fat content.csv (replication and average fat on MCV encapsulation).
▪ Fat_data for graphs with GraphPad.csv (fat percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Fat_Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Fat_Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).
▪ Flavonoid content.csv (replication and average flavonoid on MCV encapsulation).
▪ Flavonoid_data for graphs with GraphPad.csv (flavonoid percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Flavonoid_Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Flavonoid_Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).
▪ Moisture Content.csv (replication and average moisture on MCV encapsulation).
▪ Moisture_data for graphs with GraphPad.csv (moisture percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Moisture_Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Moisture_Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).
▪ Phenols content.csv (replication and average phenols on MCV encapsulation).
▪ Phenols_data for graphs with GraphPad.csv (phenols percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Phenols_Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Phenols_Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).
▪ Protein content.csv (replication and average protein on MCV encapsulation).
▪ Protein_data for graphs with GraphPad.csv (protein percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Protein_Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Protein_Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).
▪ Tannin content.csv (replication and average tannin on MCV encapsulation).
▪ Tannin_data for graphs with GraphPad.csv (tannin percentage data on MCV encapsulation for graphing with GraphPad Prism).
▪ Tannin_Ordinary one-way ANOVA Results with GraphPad.csv (results of data analysis with ANOVA using GraphPad Prism).
▪ Tannin_Multiple comparisons with GraphPad.csv (data analysis with Tukey’s multiple comparisons test using Graphpad Prism).

Data are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication).

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Author details Author details

¹ Department of Agricultural Product Technology, University of Mulawarman, Samarinda, East Kalimantan, 75119, Indonesia
² Research Center for Drugs and Consmetics from Tropical Rain Forest (PUI-PT Oktal), University of Mulawarman, Samarinda, 75123, Indonesia
³ Agricultural Sciences Doctoral Program, University of Mulawarman, Samarinda, East Kalimantan, 75123, Indonesia

Anton Rahmadi
Roles: Funding Acquisition, Investigation, Project Administration, Supervision, Writing – Review & Editing

Siti Nurjannah
Roles: Formal Analysis

Yulian Andriyani
Roles: Data Curation, Validation

Maghfirotin Marta Banin
Roles: Methodology, Project Administration

Miftakhur Rohmah
Roles: Conceptualization, Investigation, Methodology, Writing – Original Draft Preparation

Nur Amaliah
Roles: Investigation, Methodology

Kartika Sari
Roles: Data Curation, Resources

Aswita Emmawati
Roles: Data Curation, Supervision, Writing – Original Draft Preparation

Competing interests

No competing interests were disclosed.

Grant information

This work was supported by the Indonesian Ministry of Education, Culture, Research, and Technology, under grant no.: 598/UN17.L1/PG/2021, awarded to Anton Rahmadi.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Rahmadi A, Nurjannah S, Andriyani Y et al. Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (Artocarpus champeden) vinegar prepared with maltodextrin and chitosan as wall materials [version 1; peer review: 1 approved with reservations] F1000Research 2022, 11:865 (https://doi.org/10.12688/f1000research.109612.1)

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Reviewer Report 03 May 2023

Gülay Baysal, Nutrition and Dietetics, Faculty of Healthy Sciences, Istanbul Aydin Universitesi, Istanbul, İstanbul, Turkey

Approved with Reservations

https://doi.org/10.5256/f1000research.121132.r171418

This manuscript is research on “Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (Artocarpus champeden) vinegar prepared with maltodextrin and chitosan as wall materials”. I would like to say that the research is unacceptable without minor revision. ... Continue reading

This manuscript is research on “Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (Artocarpus champeden) vinegar prepared with maltodextrin and chitosan as wall materials”. I would like to say that the research is unacceptable without minor revision. Comments are as follows:

The authors should add the pre-treatments in the ash analysis to the first part of the experimental phase.
Please explain in detail all auxiliary chemicals used in the analysis by specifying their specific properties in the material section.
Number all of the formulas used in protein, fat and carbohydrate analysis and define references for each one separately.
Please discuss and interpret the following sentence in detail:
“Minerals are not significantly affected by chemical and physical treatments during processing.⁴⁵ For example, specific mineral ions can be oxidized in the presence of oxygen, but their final concentrations were not significantly affected..."
Please specify the encapsulation efficiency ratio in the following sentence:

“Encapsulation efficiency is calculated based on the number of polyphenols encapsulated."
Minor grammatical errors in English need to be corrected.
In the introduction section, add a brief analysis of the research findings on chitosan and its encapsulation as a paragraph. The following 2 articles will help you: Baysal, Olcay & Günneç (2023)¹ & Baysal et al., (2022)².

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

References

1. Baysal G, Olcay H, Günneç Ç: Encapsulation and antibacterial studies of goji berry and garlic extract in the biodegradable chitosan. Journal of Bioactive and Compatible Polymers. 2023; 38 (3): 209-219 Publisher Full Text
2. Baysal G, Olcay HS, Keresteci B, Özpinar H: The antioxidant and antibacterial properties of chitosan encapsulated with the bee pollen and the apple cider vinegar.J Biomater Sci Polym Ed. 2022; 33 (8): 995-1011 PubMed Abstract | Publisher Full Text

Competing Interests: No competing interests were disclosed.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

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Gülay Baysal, Istanbul Aydin Universitesi, Istanbul, Turkey

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Reviewer Report

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03 May 2023 | for Version 1

Gülay Baysal, Nutrition and Dietetics, Faculty of Healthy Sciences, Istanbul Aydin Universitesi, Istanbul, İstanbul, Turkey

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Approved With Reservations

This manuscript is research on “Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (Artocarpus champeden) vinegar prepared with maltodextrin and chitosan as wall materials”. I would like to say that the research is unacceptable without minor revision. Comments are as follows:

The authors should add the pre-treatments in the ash analysis to the first part of the experimental phase.
Please explain in detail all auxiliary chemicals used in the analysis by specifying their specific properties in the material section.
Number all of the formulas used in protein, fat and carbohydrate analysis and define references for each one separately.
Please discuss and interpret the following sentence in detail:
“Minerals are not significantly affected by chemical and physical treatments during processing.⁴⁵ For example, specific mineral ions can be oxidized in the presence of oxygen, but their final concentrations were not significantly affected..."
Please specify the encapsulation efficiency ratio in the following sentence:

“Encapsulation efficiency is calculated based on the number of polyphenols encapsulated."
Minor grammatical errors in English need to be corrected.
In the introduction section, add a brief analysis of the research findings on chitosan and its encapsulation as a paragraph. The following 2 articles will help you: Baysal, Olcay & Günneç (2023)¹ & Baysal et al., (2022)².

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

References

1. Baysal G, Olcay H, Günneç Ç: Encapsulation and antibacterial studies of goji berry and garlic extract in the biodegradable chitosan. Journal of Bioactive and Compatible Polymers. 2023; 38 (3): 209-219 Publisher Full Text
2. Baysal G, Olcay HS, Keresteci B, Özpinar H: The antioxidant and antibacterial properties of chitosan encapsulated with the bee pollen and the apple cider vinegar.J Biomater Sci Polym Ed. 2022; 33 (8): 995-1011 PubMed Abstract | Publisher Full Text

Competing Interests

No competing interests were disclosed.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

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Proximate analysis of the high phytochemical activity of encapsulated Mandai cempedak (Artocarpus champeden) vinegar prepared with maltodextrin and chitosan as wall materials

Abstract

Keywords

Introduction

Methods

Mandai Cempedak Vinegar (MCV)

MCV encapsulation

Moisture content analysis

Ash content analysis

Protein content analysis

Fat content analysis

Carbohydrate content by difference

Total flavonoid

Total phenol

Total tannin

Statistical analysis

Table 1. Mandai cempedak vinegar encapsulation treatment.

Result and discussion

Moisture content

Table 2. Proximate values of encapsulated mandai cempedak vinegar product.

Ash content

Protein content

Fat content

Carbohydrate content

Total flavonoid

Table 3. Phytochemical values of encapsulated mandai cempedak vinegar.

Total phenol

Total tannin

Conclusions

Data availability

Underlying data

References

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